Chapter 6.

Microbial growth

Cells contain macromolecules including DNA, RNA, proteins, lipids, and

polysaccharides. Many combinations of these molecules make up structures
like chromosome, cell wall, capsule, etc.
To divide and grow living cells should make all requirements to make cell
components.

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 Microbial Nutritional Requirements

Factors that affect microbial growth:

1. Availability of nutrients
nutrients serve as energy source as well as sources of carbon, hydrogen, oxygen, sulfur,
phosphorus, and nitrogen.
Special elements (potassium, calcium, iron, manganese, copper, zinc) and specific vitamin or
organic substances are needed by some bacteria.

2. Moisture

•Water is essential for life, as we know it. Cells are composed of between 70% and 95%
water.
•All living organisms require water to carry out their normal metabolic processes.
•Most of vegetative cells will die in very low moisture environment except dry condition
(desiccation); only endospores and protozoan cyst can survive.
•Those structures (spores) once they placed in a moist, nutrient-rich-environment, they will
grow again and produce normally

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3. pH
it’s the hydrogen ion concentration of a solution, thus, its define the acidity or alkalinity
od the solution.
Most microbes prefer neutral environment or slightly alkaline growth medium (pH 7.0-
7.4).

Acidophilic microbes prefer acidic environment like those who lives in stomach, in
pickled food or in hydrothermal vents producing sulphurous gases with pH range (2-5).
Fungi prefer acidic environment with growth medium pH range (3-6).

Alkhiphiles prefer an alkaline environment (pH  8.5), such as microbes found in

intestine, in soils laden with carbonate and Vibrio cholerae-bacteria that cause Cholera
disease- is the only human pathogen that grows well above pH 8.

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Microbes that actually prefer salty environments (such as the
concentrated salt water found in the Great Salt Lake and solar
salt evaporation ponds) are called halophilic, halo referring to
“salt” and philic meaning “to love.” Microbes that live in the
ocean, such as V. cholerae.

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5. Oxygen requirement. Microorganisms vary in their oxygen requirement.
Types of bacteria based on oxygen requirement
Type Oxygen Requirement Examples

Obligate aerobes 20-21% oxygen Mycobacteria

Microaerophilic Live in lower oxygen Neisseria gonorrhoeae

aerobes concentration (5%) Campylobacter bacteria
(cause diarrhea)
Facultative anaerobes Live in the presence or Most streptococci
absence of oxygen Staphylococci
Enterobacterioceae
Aerotolerant anaerobes Do not require oxygen for Lactobacilli
growth, but can tolerate the
presence of oxygen
Obligate anaerobes Live in the absence of Clostridium
oxygen. Die in the presence Bacteriodes
of oxygen. Why?
Capnophiles Grow in low oxygen, but Neisseria
grow better in increased Campylobacter
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carbon dioxide (5-10%) haemophilus
6.Temperature

Psychrophils (cold-loving). Optimum temperature 10 – 20 oC

Psychrotrophs. Grow at refrigerator temperature
Mesophiles (grow best at moderate temperature)
Optimum temp. 20-40 oC, most human pathogens are mesophilic because
they grow best at normal body temperature (37 oC)
Thermophiles (heat-loving). Optimum temperature greater than 50- 60 oC

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Every microorganism has an optimal, minimum, and maximum
growth temperatures.
Minimum temperature: lowest temperature that permits a
microbe’s growth and metabolism
Maximum temperature : highest temperature that permits a
microbe’s growth & metabolism
Optimum temperature : promotes the fastest rate of growth and
metabolism

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Microbial Growth: An increase in the number (not size) of cells
Binary fission: Division of an individual cell into two new cells.
Generation time : The time required for the cell population to double.
The generation time (GT) varies from one bacterial species to another.
Examples: Clostridium spp. GT = 10 min; Staphylococcus spp. GT = 20 min
((called rapid growers)
Mycobacterium tuberculosis (TB) GT = 18-24 hours (called slow growers)
Aseptic techniques are measures taken to ensure that living pathogens are
absent.
Aseptic technique is practiced in the microbiology laboratory to:
- Prevent infection of individuals
- Prevent contamination of the work environment, clinical specimen
and cultures
Pure culture: is a culture that contains only one species of organism.
A colony: is a visible mass of microbial cells that theoretically arise from one 10
cell.
 BACTERIAL GRWOTH CURVE
The growth curve of particular species of bacterium determined by
growing a pure culture of the organisms in a liquid medium. Samples
collected at fixed intervals (e.g., every 30 minutes), number of viable cells is
determined.
1. Lag phase: Bacteria absorb nutrients, synthesize enzymes and prepare for cell
division.
No increase in the number of bacteria in the Lag phase.
2. Logarithmic growth phase: Bacteria multiply rapidly, maximum growth continues
as long as cells have adequate nutrients & favorable environments.
The growth rate is the greatest during the Log phase
3. Stationary phase: Less nutrients and more toxic waste products, the number of
dividing cells equals the number of dying cells. The rate of division slows.
4. Death phase: bacteria die at a rapid rate. If bacteria is a spore-former, it will
produce spores to survive beyond this phase. Bacteria may undergo morphological
changes.

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 The chemostat (continuous culture):
Many industrial and research procedures depend in
the maintenance of an essential species of microorganism.
These are continuously cultured in in a controlled
environment called “Chemostat” which regulates the
supply of nutrients and the removal of waste products

How to measure microbial growth

Direct Methods
A) Total microscopic count
Disadvantages:
1) dead cells are not distinguished from living cells;
2) small cells are difficult to see under the microscope;
3) precision is difficult to achieve;
4) we need a phase contrast microscope;
5) not a good method for cell suspensions of low density.
Indirect Methods
Turbidity: Bacteria is suspended in in a
liquid and a small amount is put in glass
container which is placed in a light
device called spectrophotometer

2- Viable (colony) count

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Culturing of bacteria
Different types of culture media are used depending on the type of
bacteria As bacteria grow, they produce many waste products and
secretions, some of which are enzymes enable them to invade their hosts
and cause disease. Biochemical activities help in the identification of
certain types of bacteria in the laboratory.

Culturing of obligate intracellular pathogens in the laboratory:

Viruses = can be propagated using cell cultures
Rickettsias and Chlamydias = can be propagated using embryonated
chicken eggs or laboratory animals .

Culturing of fungi in the laboratory:

Fungi (yeasts, molds, and dimorphic fungi) will grow in various media
An example of media is “Sabouraud Dextrose Agar (SDA)”.
Care should be taken to avoid inhaling fungal spores (some are infectious).

Culturing of protozoa in the laboratory:

Most clinical microbiology laboratory do not culture protozoa but
techniques are available to for culturing some protozoa.
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In general, they use microscopy to see cysts or trophozoites in laboratories